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Sept. (Ascom Ycetes) Rijksherbarium, Leiden Pyronemataceae (Merkus PERSOONIA Published by the Rijksherbarium, Leiden Volume Part 8, 3, pp. 227-247 (1975) Ultrastructure of the ascospore wall in Pezizales (Ascomycetes) —III. Otideaceae and Pezizaceae Emily Merkus Rijksherbarium, Leiden (With Plates 34-45) The of wall and ornamentation of is studied development layers ascospores with the electron microscope in members of the Otideaceae and Pezizaceae. and the in Ascodesmis Primary wall, endospore, epispore developin sameway as and the Pyronemataceae; the development of the secondary wall and the formation of the patterns ofornamentation resemble that in the Pyronemata- attention is ceae. Special paid to specialized plasmic structures. Introduction earlier I of In papers (Merkus, 1973, 1974) reported my electron microscopy the ultrastructure ofthe wall in Pezizales and discussed theresults Ascodesmis ascospore on and the Pyronemataceae sensu Eckblad (1968). The continuationof this and the results in present paper represents a study gives the Otideaceae and which identical species belonging to Pezizaceae, are practically with the Otideae and Aleurieae ofthe Aleuriaceae of Le Gal (1947: 284). REVIEW OF EARLIER WORK brief review is Like in my paper on the Pyronemataceae (Merkus, 1974), a given of Gal's of ornamentation of the of Le light microscopy the patterns ascospores the Pezizaceae (1947). Though this involves duplication of some of the data, it gives a better overall insight and makes it easier to interpret new results. According to the rules of the international code of botanicalnomenclature some of the names used by Le Gal (see footnotes) had to be changed. ornamentation. The species of Pezizaceae studied by Le Gal develop simple spore In all the wall arises and it is the species a primary ornamentation on present; orna- mentation consists of callose and pectine formations and is of sporal origin. Persoonia, Vol. 8, Part 2 was issued 12 Sept. 1975 227 PERSOONIA Vol. Part 228 8, 3, 1975 The primary wall is covered by an "assise sous-perisporique" and a "pellicule membranaire". The "assise sous-perisporique" is formed before ornamentation develops. The "pellicule membranaire" is termed "coque interperisporique" ifit is formed at the same time as the ornamentation and is penetrated by the substance of the the and the substance of the ornamentation; "coque interperisporique" orna- mentation both grow into one, the "coque interperisporique" also consisting of callose and pectine. The "pellicule membranaire" is termed "tunique externe de 1'assise" ifit is formed before ornamentation develops and is not penetrated by the substance of the ornamentation. Pulparia persoonii (Crouan) Korf & al. apud Korf 1 develops simple ornamentation that is formed between the primary wall and its covering layers. 2 P. badia Pers. 3 P. P. 4 P. Peziza succosa Berk., , per Mérat, . eckinospora Karst., trachycarpa 5 6 7 Curr., P. apiculata Cooke, and P. reperta (Boud.) Moser develop simple ornamen- which of the tation, the substance of penetrates the covering layers primary wall and is the the of the deposited on "coque interperisporique". During development ornamentation is the outside of the In the a "perispore" present on ascospores. first four species the "perispore" disappears in a later stage; in the other two species it remains. Le Gal does of the ornamentation ofthe not give a description patterns ascospores of the Otideaceae. of attention Though the ascoplasm is not the main subject thepresent study special is paid to it. It reveals the development of globular structures, like oil bodies and other specialized globules of a still unknown substance; these were also described by Guilliermond (1904, 1910, 1920), the latter as "corpuscules metachromatiques". The development of the structures and their relation to ascospore development is discussed. MATERIALS AND METHODS collected in the The material of the species in the present study was Netherlands, and in the list data about the in France, Germany; following gives some specimens and their localities: Otidea alutacea (Pers. per S. F. Gray) Mass. - van Brummelen needles of "foret de 73-501, on Picea, Liciat", Oyonnax, Ain, France, 7.X.1973 — soil The (L); Piepenbroek 837, on under Quercus, t Joppe, Gorssel, Gelderland, O. Boud. — Brummelen the Netherlands, 12.X.1974 (L) ; bufonia (Pers.) van 4073, on ground under Betula, Schoorl, North Holland, The Netherlands, 3.IX. 1973 (L); — 1 Marcelleina Brumm. Plicaria persoonii (Crouan) Boud.; syn. persoonii (Crouan) 2 Galactinia succosa (Berk.) Sacc. badia 2 Galactinia (Pers. per Merat) Boud. 4 Aleuria umbrina (Boud. apud Cooke) Gill.; not Peziza umbrina Pers. 2 Plicaria trachycarpa (Curr.) Boud. 6 Aleuria apiculata (Cooke) Boud. 7 Aleuria reperta (Boud.) Boud. MERKUS: Ultrastructure wall in and of ascospore Otideaceae Pezizaceae 229 under 't Piepenbroek 802, on soil, among grasses Quercus, Joppe, Gorssel, Gelderland, The O. onotica S. F. Fckl. — Netherlands, 28.VII.1974 (L) ; (Pers. per Gray) van the Brummelen 4638, on ground under oaks, Koningshof, Overveen, Bloemendaal, North The 2.XI. Dur. Lév. Holland, Netherlands, 1974 (L) ; Peziza ammophila & apud Dur. — collected during a field trip of the Dutch Mycological Society, sandy The X. dunes, Hollumerduinen, Ameland, Friesland, Netherlands, 27. 1973 (L) ; Pers. Pers. — Brummelen dead culms of P. ampliata per van 4074, on Phragmites, Nederhorst den Berg, North Holland, The Netherlands, 12.V.1973 (L); P. badia Pers. per Merat — Piepenbroek 806, on sandy soil, near estate "Dörth", Gorssel, — Gelderland, The Netherlands, 28.VII.1974 (L) ; Piepenbroek 811, on sandy soil, estate "Ampsen", Lochern, Gelderland, The Netherlands, 10.VIII.1974 (L); P. Dennis — badiofusca (Boud.) Piepenbroek 746, on clay soil, Duursche Waarden between The P. emileia Cooke — Olst and Wijhe, Overijsel, Netherlands, 23.VI. 1974 (L) ; Piepenbroek 556, on burnt soil, "de Bannink", Colmschate, Overijsel, The Netherlands, Dennis — 3.VI. 1973 (L); P. michelii (Boud.) van Brummelen 4500, on sandy soil, Waterdijk, Diepenveen, Overijsel, The Netherlands, 15.VII. 1974 (L) ; P. petersii Berk. — Siteur, on burnt soil, Eindhoven, North Brabant, The Netherlands, 21.VII. Nannf. in Lundell — 1974 (L); P. plebeia (Le Gal) & Nannfeldt Piepenbroek 773, on The VII. sandy soil, Epserbos, Epse, Gelderland, Netherlands, 21. 1974 (L); P. Bres. — Brummelen burnt 't praetervisa van 4072, on soil, Joppe, Gorssel, Gelderland, The Netherlands, 6.VIII.1973 (L); — Piepenbroek 734, on burnt soil, "Klein Noor- The dijk", Wilp, Voorst, Gelderland, Netherlands, 27.VII.1974 (L); P. succosa Berk. — Brummelen soil under Waarden between van 431 /, on Salix, Hengforder Olst and The P. succosella Le Gal & Wijhe, Overijsel, Netherlands, 15.VII.1974 (L) ; — Brummelen & soil Romagn. van Piepenbroek 4510, on under Salix, Waterdijk, The P. Curr. — Diepenveen, Overijsel, Netherlands, 15.VII. 1974 (L) ; trachycarpa Piepenbroek 807, on soil among mosses, near estate "Dörth", Gorssel, Gelderland, The Netherlands, 28.VII. 1974 (L) ; P. vesiculosa Bull, per St-Am. — van Brummelen 4080, on soil in hot-house, Kortenhoef, North Holland, The Netherlands, 28.1.1974 (L); Pulparia persoonii Korf & al. apud Korf — Piepenbroek 575b, on loamy soil, Duursche Waarden near Fortmond, Overijsel, The Netherlands, 19.VIII. 1973 (L) ; — Piepen- broek 738, on loamy soil, Duursche Waarden between Olst and Wijhe, Overijsel, The Netherlands, 23.VI.1974 (L); van Brummelen 4313, on loamy soil, Duursche Waarden between Olst and Wijhe, Overijsel, The Netherlands, 15.VII. 1974 (L); Pustularia Fckl. — cupularis (L. per Fr.) Piepenbroek 772, on sandy soil, Epserbos, Epse, Gelderland, The Netherlands, 21.VII. 1974 (L); Sowerbyella radiculata (Sow. per Fr.) Nannf. — Haas, under Picea, Kreis Heidenheim, Schwabisch Alb, Germany, — from X. 4.X. 1973 (L); exposition, Oyonnax, Ain, France, 22. 1973 (L). After collection fromtheir substratum in the field, the apothecia were placed in the fixative. Further treatment of the apothecia followed much the same procedure as described in former studies but a few additional remarks are called for. In addition to the former used, KMn0 or percentages 1% 4 1% glutaraldehyde was applied as primary fixative before postfixation with OSO4 to improve results. PERSOONIA Part 230 Vol. 8, 3, 1975 In order to solve the problem of inadequate impregnation of the embedding medium in the relatively hard material, in some cases the usual Epon embedding medium according to Luft ( 196 1) was replaced by a low-viscosity embedding medium according to Spurr (1961), with the additive dibutyl phtalate (Clemen$on, 1973). The material was transferred to the Spurr embedding medium via an ethanol series, and mixtures of the 1 00% ethanol, 100% acetone, acetone and Spurr embedding medium; polymerization lasted 12 hours at 60 °C. The of the used of components Spurr embedding medium were at a rate 10.0 g 26.0 ERL-4206, 6.0 g D.E.R. 736, g nonenylsuccinic anhydride, 0.4 g dimethyla- minoethanoland 0.8 g dibutyl phtalate. All sections were cut with a diamond knife. OBSERVATIONS The species studied so far all showed practically the same ultrastructure and a general description could be given. ofthe data reveals thatthe ultrastructure of ofthe Otidea- A review present species ceae and Pezizaceae answers to this description. This holds particularly for the of the delimitationof in the younger stages development; ascospores ascoplasm by unit the their distri- two delimiting membranes, form and structure of organels and bution in epiplasm and sporoplasm, and the development of
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